REPRODUCTIVE BIOLOGY OF DIPLOID AND TRIPLOID APIOS AMERICANA (LEGUMINOSAE)

Apios americana Medicus (Leguminosae; Phaseoleae), a tuberous, herbaceous vine, consists of diploid and triploid clones which are morphologically nearly identical. Although A. americana is widespread in eastern North America and flowers profusely throughout its range, fruit set is low. In part, this is because populations north of Connecticut consist almost exclusively of sterile triploid individuals. Although triploid populations occur south of this region, diploids predominate. Fruit set is also low in diploids. We attribute this to partial self-incompatibility and to a low rate of floral visits by leaf-cutter bees (Megachile spp.), the only legitimate pollinator. Visitation is easily measured because flowers have a nonrepetitive explosive tripping mechanism which must be triggered by insects. In two Connecticut populations only 10% of 299 flowers were tripped, and on average only about one-half of these set fruit. We conclude that A. americana maintains populations by vegetative propagation, but expands its range and retains variability through sexual reproduction of diploids.     

RESTRICTION FRAGMENT ANALYSIS OF CHLOROPLAST DNA AND THE SYSTEMATICS OF VIGUIERA AND RELATED GENERA (ASTERACEAE: HELIANTHEAE)

Phylogenetic relationships among 31 species representing variously Helianthus, Helianthopsis, Heliomeris, Simsia, Viguiera, and Tithonia were assessed by chloroplast DNA restriction site mapping. A total of 147 mutations including 92 informative ones was detected using 16 restriction enzymes, with an estimated sequence divergence within the group of 1.4%. Parsimony analysis produced a single most parsimonious tree with a length of 161 steps and a consistency index of 0.91. Statistically significant clades, as assessed by the bootstrap method, correspond to a number of taxa recognized previously, including Helianthus (3 species), Helianthopsis (5 species), and several groups within Viguiera, including sect. Maculatae (4 species), the Baja California group (6 species), sect. Paradosa (2 species) and V. dentata (3 samples). However, species of Viguiera collectively form a highly paraphyletic group relative to species of other genera. Helianthus and Helianthopsis were separated into different clades, supporting their recent segregation. Placement of H. porteri in Helianthus rather than Heliomeris was confirmed; the single sample of the latter genus was most similar to the Baja California group of Viguiera. An expected relationship between Simsia (2 species) and one member of Viguiera ser. Grammatoglossae was confirmed (although not with two other putatively related members of Viguiera) and an unexpected relationship between Simsia and Tithonia was suggested. The presence of Mexican taxa as the more basal groups in the tree points toward a possible Mexican origin for Viguiera and related genera. A molecular clock hypothesis is rejected in many pairwise comparisons involving woody taxa with herbaceous ones, although it could not be rejected in most pairwise comparisons involving taxa of similar habit.     

CHROMOSOME RELATIONSHIPS OF DIPLOID SPECIES OF GRINDELIA (COMPOSITAE) FROM COLORADO,NEW MEXICO,AND ADJACENT AREAS

Previous studies of chromosome relationships of Grindelia species recognized three basic genomes designated Oxylepis, Hallii, and Havardii. Differences are based on different end arrangements of the chromosomes resulting from reciprocal translocations. This report will review and give additional information about the genomes and interrelationships of 17 species. All of the species are diploids (2n = 12) and show six bivalents at meiosis. Species in this study that have the Oxylepis genome are G. oxylepis var. eligulata, G. fastigiata, G. inornata, G. revoluta, and G. squarrosa. Species that have the Havardii genome included G. havardii, G. grandiflora, G. lanceolata, G. littoralis, and G. texana. The Hallii genome is present in G. camporum var. davyi and G. procera. Hybrids of species with the same genome have six bivalents at meiosis. Hybrids between species with the Oxylepis genome and those having the Havardii genome have four bivalents and one quadrivalent at meiosis. Likewise for Oxylepis x Hallii hybrids. A new genome is presented for G. subalpina which would explain the configurations of two bivalents and two quadrivalents observed in G. subalpina x G. havardii and G. subalpina x G. fastigiata hybrids. This is designated the Subalpina genome. Species tested but with genomes as yet undetermined are G. acutifolia, G. arizonica, G. nana, and G. scabra.     

CYTOHISTOLOGICAL STUDIES OF CYTOCHIMERIC AND TETRAPLOID GRAPES

Thompson, M. M., and H. P. Olmo. (U. California, Davis.) Cytohistological studies of cytochimeric and tetraploid grapes. Amer. Jour. Bot. 50(9): 901–906. Illus. 1963.—Cytohistological studies were made on 62 different grape clones suspected to be tetraploids to determine if they were chimeras, and if so, what type. The chromosomal constitution of the apical layers in the shoot was determined on the basis of cell and nucleus size, size of chromosome mass, and chromosome estimates or counts. Three cytological types were observed; 2–4,² 4–2, and 4–4. The possible types of chimeras and their stability depend upon the organization of the shoot apex. In most buds there appeared to be a biseriate tunica covering a corpus. However, the number of tunica layers fluctuated from 1 to 3, and there were occasional periclinal divisions in L-II. This lack of discreteness of the second layer probably accounts for the homogeneity of all tissue beneath the superficial layer. The rarity of periclinal divisions in L-I explains the stability of 2–4 types, but reversions are not impossible. The relative frequency of the different types of sports is discussed in relation to their possible origin. The theory of intraclonal variability developed in grapes by Reichardt (1955) and Rives (1961) on the basis of a multilayered shoot apex is re-evaluated under the concept of a functionally uniseriate tunica.     

THE TAXONOMIC SIGNIFICANCE OF ANTHOCHLORS IN THE SUBTRIBE COREOPSIDINAE (COMPOSITAE,HELIANTHEAE)

Anthochlors (chalcones and aurones) are found in a number of plant families including the Compositae. Within this family they were once thought to occur only in the subtribe Coreopsidinae of the tribe Heliantheae. More recent studies show them to occur also in the tribes Cardueae, Eupatorieae, Helineae, Inuleae, and Lactuceae. This has suggested that anthochlors are no longer good taxonomic markers for the Coreopsidinae. A survey of 69 of approximately 210 genera of the Heliantheae shows anthochlors present only in the Coreopsidinae except for Helianthus, Simsia, Tithonia, and Viguiera, closely related genera of the subtribe Helianthinae. Of the 32 genera of the Coreopsidinae recently recognized, 30 were surveyed from available material and all contain anthochlors except Guardiola and Venegasia. The results indicate that, despite some variation, anthochlors are still good taxonomic markers for the Coreopsidinae. This represents the only case within the family in which a particular type of flavonoid is taxonomically diagnostic at the subtribal level.     

CHLOROPLASTS OF MONOPLOID AND DIPLOID OENOTHERA HOOKERI

Comparison of leaves from diploid Oenothera hookeri plants with leaves of their monoploid derivatives revealed the following differences: the leaves of the diploids are almost twice as thick as those of the monoploids; the diploids have only one-fifth as many cells per unit leaf area as do the monoploids; leaf tissue of the diploids has an average of 36.3 chloroplasts per cell compared with 27.4 for the monoploids; chloroplasts of the diploids average 4.68 μm in length, those of the monoploids, 3.74 μm; chloroplasts from the diploids contain about seven times more chlorophyll than those of the monoploids and have an average of seven thylakoids per granum compared with an average of only three for the monoploids; the average chlorophyll a/b ratio for the diploid chloroplasts is 2.98 compared with 3.48 for the monoploids.     

CHROMOSOME NUMBERS AND THEIR SYSTEMATIC IMPLICATIONS IN LIPOCHAETA (COMPOSITAE:HELIANTHEAE)

Chromosome counts are reported for 71 collections in 14 taxa of Lipochaeta DC. (Compositae) of the Hawaiian Islands. Counts for 13 of the taxa are first reports. Lipochaeta contains diploids, n = 15, and tetraploids, n = 26. Ploidy level is taxonomically significant for recognizing sections, and on the basis of these data, together with morphological and chemical evidence, major realignments of species into the secions of Lipochaeta have been made.     

EVIDENCE FOR A CLOSE RELATIONSHIP BETWEEN IOSTEPHANE AND VIGUIERA (ASTERACEAE: HELIANTHEAE)

The phylogenetic relationship of Iostephane is assessed using data from morphology, flavonoid chemistry, and chloroplast DNA and nuclear ribosomal DNA restriction fragment analysis. Morphological evidence supports placement of Iostephane in subtribe Helianthinae, but fails to clarify the placement of the genus within this assemblage. Further evidence for the placement of Iostephane in subtribe Helianthinae is provided by the presence in all species of the genus of floral flavonoids of the chalcone/aurone type, which provides a distinctive trait for the subtribe within the tribe Heliantheae. Analysis of chloroplast DNA from two species of Iostephane, I. heterophylla and I. madrensis, in comparison to Viguiera and related genera indicates that the restriction site patterns with 16 enzymes for the Iostephane species are virtually identical to one another as well as to those of Viguiera sect. Maculatae. Data from restriction fragment patterns of nuclear rDNA are concordant with the results from chloroplast DNA in suggesting a direct relationship between the two groups. The close phylogenetic relationship between Iostephane and Viguiera sect. Maculatae suggested by the DNA restriction fragment data was not suggested by any other set of data.     

A NEW SPECIES OF COREOCARPUS (COMPOSITAE: HELIANTHEAE) FROM SAN PEDRO NOLASCO ISLAND,MEXICO

A new species of Coreocarpus endemic to San Pedro Nolasco Island, Mexico, C. sanpedroensis , is described and illustrated.     

CHLOROPLAST-DNA AND ALLOZYMIC VARIATION IN DIPLOID AND AUTOTETRAPLOID HEUCHERA GROSSULARIIFOLIA (SAXIFRAGACEAE)

Diploid and autotetraploid populations of Heuchera grossulariifolia occur throughout mountainous regions of the Pacific Northwest. Controlled greenhouse crosses indicated that the two cytotypes are largely reproductively isolated. Fourteen diploid and 11 tetraploid populations were analyzed electrophoretically. Individual tetraploid plants expressed up to four alleles per isozyme locus, and tetraploid populations had significantly higher levels of heterozygosity than diploids. Mean observed heterozygosity was 0.159 for tetraploid populations and 0.058 for diploid populations. The patterns of allelic distribution between cytotypes suggested multiple origins of autotetraploids. This hypothesis was supported by restriction-site analysis of chloroplast-DNA (cpDNA) variation which indicated that there had been at least three independent origins of tetraploids. Electrophoretic data, in conjunction with a cpDNA-based phylogeny and geographic distribution of populations, suggest that autopolyploid populations evolved several times as migration of diploids occurred down river systems. This study further supports the contention that autopolyploidy can be a common and successful speciation process in some groups of plants.     

EVOLUTION OF CYTOPLASMIC FACTORS IN TETRAPLOID CULTIVATED POTATOES (SOLANACEAE)

Analyses were made, by study of reciprocal crosses, of cytoplasmic factors of Solanum tuberosum ssp. andigena and S. tuberosum ssp. tuberosum including both northern hemipshere cultivated clones and clones from Chiloe Archipelago in Coastal Chile. The purpose was to compare the cytoplasmic factors of these three sorts of S. tuberosum with those of their diploid ancestors and with one another as aids in determining their positions on pathways of evolutionary descent. Preliminary to making the comparisons it was shown that the cytoplasm-chromosomal gene interaction that produces deformed flowers in diploid forms of Solanum functions also in induced autotetraploids, with deformed flowers being expressed in [df⁸]-dfdfdfdf individuals. Plants of the S. stenotomum-phureja complex, putative ancestors of S. tuberosum ssp. andigena, were known from earlier work to have the [df⁸] plasmon factor, but ssp. andigena itself was found to lack it. A newly discovered cytoplasmic abnormality, anther-lobe overlap, is expressed in plants having plasmon factor [lo⁸] and homozygous for recessive alleles lo. The [lo⁸] plasmon factor is present in S. phureja and in S. tuberosum ssp. andigena. Altogether, ssp. andigena potatoes match their putative stenotomum-phureja ancestors in eight of the nine plasmon factors identified to date, differing with them only in the lack of [df⁸]. Northern hemisphere and Chiloe clones of S. tuberosum ssp. tuberosum lack the [df⁸] and the [lo⁸] plasmon factors. Chiloe clones are identical to northern hemisphere cultivated ones in all eight plasmon factors for which they were tested. Both contrasted with ssp. andigena, their putative ancestor, in eight of nine factors tested, resembling it only in their common lack of [df⁸]. The evidence supports the idea that clones cultivated now in the northern hemisphere are descendants of direct imports from coastal Chile. The clone Rough Purple Chili was an example of an imported clone that became important in potato breeding, but it has not been clear whether it was brought from Chile or from Andean regions of northern South America. The leaf form and cytoplasmic factors of Garnet Chili, a first generation derivative of Rough Purple Chili, were found to be those of S. tuberosum ssp. tuberosum. Rough Purple Chili is, therefore, an example of such direct importation from Chile.     

THE CYTOLOGY AND PHYLOGENETICS OF THE DIPLOID SPECIES OF GOSSYPIUM

Meiotic chromosome behavior of 11 inter-genomic hybrids of Gossypium (2n = 26) were investigated. Per cell univalent frequencies at meiotic metaphase I in these hybrids were: A genome × Cgenome—G. herbaceum × sturtianum, 10.53; G. herbaceum × australe, 18.05. A genome × E genome—G. smnalense × arboreum, 21.82. B genome × C genome—G. anomalum × sturtianum, 9.23; G. anomalum × australe, 13.11. B genome × D genome—G. anomalum × klotzschianum, 17.45; G. anomalum × raimondii, 18.83. C genome × D genome—G. robinsonii × davidsonii, 12.77; G. sturtianum (armourianum × thurberi), 8.63. C genome × E genome—G. somalense × australe, 23.78; G. somalense × bickii, 25.58. Trivalent and quadrivalent frequencies were relatively high for those hybrids involving a C genome species, indicating that a reciprocal translocation differentiates the C genome from the A, B, D, and E genomes. The results of this study and the data of similar studies cited from the literature on Gossypium cytogenetics are discussed relative to the phylogenetics and evolution of the major (genome) groups of Gossypium and their constituent taxa.     

GENETIC DIFFERENTIATION,SYMPATRIC SPECIATION,AND THE ORIGIN OF A DIPLOID SPECIES OF STEPHANOMERIA

Evidence is presented that a geographically peripheral population of the annual Stephanomcria exigua ssp. coronaria (Compositae), a widespread and ecologically diverse species, has recently given rise by a process of sympatric speciation to a diploid species presently designated “Malheurensis.” The new species comprises less than 250 individuals and is found only at a single locality in eastern Oregon where it grows interspersed with its parental population. Stephanomeria exigua ssp. coronaria is an obligate outcrosser and “Malheurensis” is highly self-pollinating. Reproductive isolation is maintained by differences in breeding system, a crossability barrier that reduces seed set following cross-pollination between them, and reduction in hybrid fertility caused by chromosomal structural differences. They are very similar morphologically. Electrophoretic analyses of seven enzyme systems demonstrate that all the alleles but one at the controlling 13 gene loci in “Malheurensis” are identical to alleles in ssp. coronaria. The new species displays certain maladapted features including loss of the specific requirements for seed germination characteristic of the progenitor population of ssp. coronaria. The origin of “Malheurensis” appears to be an exception to the theory of geographical speciation because spatial isolation is not necessary at any time for the origin or establishment of its reproductive isolating barriers. The nature of these barriers plus the genetic homogeneity of the species are compatible with the hypothesis that it derives from a single progenitor individual. Very little genetic change is involved initially in this mode of speciation.     

SUBCULTIVATIONS, SPLITS, DOUBLINGS, AND GENERATIONS IN CULTURES OF HUMAN DIPLOID FIBROBLASTS

Mathematical expressions are provided relating subcultivations to population doublings and population doublings to generations. Account is made for the increasing number of cells which do not attach upon subcultivation as well as the increasing number of non-dividers. The expressions reconcile published differences in longevity and reveal that the in vitro lifetime of human diploid cells measured in cell generations is far greater than previously suspected.